TARGET SPECIFIC GENE THERAPY OF PROSTATE CANCER

前列腺癌的靶向特异性基因治疗

• 批准号: 2733121
• 负责人: Shen Pang
• 金额: $ 7.44万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-07-01 至 2000-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2733121
• 关键词: Adenoviridae; SCID mouse; carcinoma; combination cancer therapy; cytomegalovirus; diphtheria toxin; disease /disorder model; gene expression; gene targeting; gene therapy; genetic promoter element; human genetic material tag; kidney neoplasms; neoplasm /cancer genetics; neoplasm /cancer therapy; neoplastic cell; nonhuman therapy evaluation; prostate neoplasms; prostate specific antigen; recombinant virus; reporter genes; thymidine kinase; tissue /cell culture; transfection /expression vector

DESCRIPTION: (Applicant's Abstract) Prostate cancer is the leading neoplasm in American men. Two hundred thousand new cases will be diagnosed in 1994 resulting in 38,000 deaths. Thus far, no effective treatment has been developed for patients with advanced, and especially hormone refractory, disease. The development of novel therapeutic strategies is paramount. The use of suicide genes to selectively eradicate tumor cells has shown promise. Driven by tumor tissue-specific promoters, these genes can be transcribed selectively in the neoplastic cells and subsequently cause their destruction. The applicant has characterized a prostate tissue-specific promoter, derived from the 5' flanking sequence of human prostate-specific antigen (PSA) gene. This promoter element has shown strong activity in PSA- producing prostate cancer cells, but not in non-PSA-producing cells. Since the tumor cells from vast majority of patients with advanced prostate disease produce PSA, utilizing this PSA promoter to drive a therapeutic gene exclusively in prostate tissue should alleviate concerns of systemic toxicity while being extremely effective. He has increased the promoter activity four- to five- fold by inserting the cytomegalovirus (CMV) enhancer upstream. Simultaneously, he has analyzed different delivery vectors to maximize gene transfer into prostate tumor cells, and found that the adenoviral vector offers significant advantages due to its high infectivity and gene expression. He will now create an adenoviral construct with a suicide effector gene driven by the CMV-PSA promoter, in the hope that this will selectively eradicate prostate tumor cells. The thymidine kinase gene (TK) has been successfully used to treat several human neoplasms. When a tumor cell transfected with TK is exposed to ganciclovir, DNA replication is disrupted and the cell is eventually destroyed. Unfortunately, prostate cancer cells replicate slowly and may not be susceptible to the actions of a therapeutic gene that requires active cell division. In this case, a suicide gene with direct lytic activity may be better. An example of this is diphtheria toxin A. Hence, the applicant will try both types of agents and assess which is most effective for prostate cancer gene therapy: the TK gene and the DTA gene, alone and in combination with TK. This proposed strategy will be tested in both cell culture and animal models. From in vitro experiments will determine the efficacy and selectivity of the chosen suicide gene. With this basic knowledge, we will then infect tumor- bearing SCID mice with our tissue-specific promoter-driven adenoviral vector to define in vivo selectivity and safety.

描述：（申请人摘要）前列腺癌是主要的癌症 美国男性的肿瘤。 新增病例将达二十万 1994年确诊，导致38,000人死亡。 到目前为止，还没有有效的 已经为晚期患者，特别是晚期患者开发了治疗方法 激素难治，疾病。 新型治疗药物的开发 战略是最重要的。 利用自杀基因选择性地 根除肿瘤细胞已显示出希望。 由肿瘤组织特异性驱动 启动子，这些基因可以在肿瘤细胞中选择性转录 细胞并随后导致其破坏。 申请人已经表征了前列腺组织特异性启动子， 源自人前列腺特异性抗原的 5' 侧翼序列 （PSA）基因。 该启动子元件在 PSA- 产生前列腺癌细胞，但不产生非 PSA 细胞。 由于肿瘤细胞来自绝大多数晚期患者 前列腺疾病会产生 PSA，利用该 PSA 启动子来驱动 专门存在于前列腺组织中的治疗基因应该可以缓解 尽管非常有效，但仍担心全身毒性。 他有 通过插入，启动子活性增加了四到五倍 巨细胞病毒 (CMV) 增强子上游。 同时，他分析了不同的传递向量以最大化 基因转移到前列腺肿瘤细胞中，发现腺病毒 载体由于其高传染性和 基因表达。 他现在将创建一个腺病毒构建体 由CMV-PSA启动子驱动的自杀效应基因，希望 这将选择性地根除前列腺肿瘤细胞。 胸苷 激酶基因（TK）已成功用于治疗多种人类疾病 肿瘤。 当转染 TK 的肿瘤细胞暴露于 更昔洛韦，DNA复制被破坏，细胞最终被破坏 被毁了。 不幸的是，前列腺癌细胞复制缓慢且 可能对需要的治疗基因的作用不敏感 细胞分裂活跃。 在这种情况下，具有直接裂解作用的自杀基因 活动可能会更好。 白喉毒素 A 就是一个例子。 因此，申请人将尝试两种类型的代理并评估哪种类型更适合 对前列腺癌最有效的基因治疗：TK基因和DTA 基因，单独或与TK组合。 该拟议战略将是 在细胞培养和动物模型中进行了测试。 来自体外 实验将确定所选药物的功效和选择性 自杀基因。 有了这些基础知识，我们就可以感染肿瘤了—— 携带我们的组织特异性启动子驱动的腺病毒的 SCID 小鼠 载体来定义体内选择性和安全性。